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Photosensitized mefloquine induces ROS-mediated DNA damage and apoptosis in keratinocytes under ambient UVB and sunlight exposure

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Abstract

The present study illustrates the photosensitizing behavior of mefloquine (MQ) in human skin keratinocytes under ambient doses of UVB and sunlight exposure. Photochemically, MQ generated reactive oxygen species superoxide radical, hydroxyl radical, and singlet oxygen through type I and type II photodynamic reactions, respectively, which caused photooxidative damage to DNA and formed localized DNA lesions cyclobutane pyrimidine dimers. Photosensitized MQ reduced the viability of keratinocytes to 25 %. Significant level of intracellular reactive oxygen species (ROS) generation was estimated through fluorescence probe DCF-H2. Increased apoptotic cells were evident through AO/EB staining and phosphatidyl serine translocation in cell membrane. Single-stranded DNA damage was marked through single-cell gel electrophoresis. Mitochondrial membrane depolarization and lysosomal destabilization were evident. Upregulation of Bax and p21 and downregulation of Bcl-2 genes and corresponding protein levels supported apoptotic cell death of keratinocyte cells. Cyclobutane pyrimidine dimers (CPDs) were confirmed through immunofluorescence. In addition, hallmarks of apoptosis and G2/M phase cell cycle arrest were confirmed through flow cytometry analysis. Our findings suggest that MQ may damage DNA and produce DNA lesions which may induce differential biological responses in the skin on brief exposure to UVB and sunlight.

Mefloquine is photosensitized by UVB and sunlight exposure at an appropriate dose and generates ROS involving both type I and type II photosensitization mechanisms. These ROS primarily damage DNA, cell membrane, and membrane-bound organelles. MQ differentially affects various biological processes which end into apoptosis of the cell

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Acknowledgments

The authors wish to thank the Director CSIR-IITR for his valuable support in this study.

Funding

This work is supported by University Grants Commission (UGC) and CSIR network project, UNDO (BSC 0103), New Delhi, India.

Conflict of interest

The authors declare that there are no conflicts of interest.

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Authors and Affiliations

  1. Photobiology Division, CSIR-Indian Institute of Toxicology Research, M.G. Marg, Lucknow, 226001, Uttar Pradesh, India

    Neera Yadav, Ashish Dwivedi, Syed Faiz Mujtaba, Ankit Verma & Ratan Singh Ray

  2. Developmental Toxicology, CSIR-Indian Institute of Toxicology Research, M.G. Marg, Lucknow, 226001, Uttar Pradesh, India

    Rajnish Chaturvedi

  3. Faculty of Pharmaceutical Sciences, College of Pharmacy, Pt. B.D.S. University of Health Sciences, Rohtak, Haryana, India

    Neera Yadav, Syed Faiz Mujtaba & Gajendra Singh

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  1. Neera Yadav
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  2. Ashish Dwivedi
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  3. Syed Faiz Mujtaba
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  4. Ankit Verma
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  5. Rajnish Chaturvedi
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  6. Ratan Singh Ray
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  7. Gajendra Singh
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Correspondence to Ratan Singh Ray.

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Yadav, N., Dwivedi, A., Mujtaba, S.F. et al. Photosensitized mefloquine induces ROS-mediated DNA damage and apoptosis in keratinocytes under ambient UVB and sunlight exposure. Cell Biol Toxicol 30, 253–268 (2014). https://doi.org/10.1007/s10565-014-9280-7

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  • DOI: https://doi.org/10.1007/s10565-014-9280-7

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